Thermostable ribonuclease H and genetic constructs therefore

Details Thermostable ribonuclease H and genetic constructs therefore Thermostable ribonuclease H and genetic constructs therefore

1995-06-05

1996-03-19

Wax, Robert A.

Chemistry: molecular biology and microbiology

Vector, per se

435199, 536 232, 536 234, C12N 922, C12N 1555, C12N 1562

Patent

active

055003702

ABSTRACT:
An essentially pure preparation of thermostable RNase H isolated from Thermus flavus is disclosed. In one preferable form of the invention, the RNase H is capable of biological activity after incubation at temperatures equal to or greater than 70.degree. C. for at least ten minutes. The present invention is also a method of digesting RNA polymers that are in duplex form with a DNA molecule. The method comprises exposing the duplex to the isolated thermostable RNase H. The present invention is also a genetic construct capable of expressing a thermostable RNase H.

REFERENCES:
patent: 3896000 (1975-07-01), Kamekura et al.
patent: 4889818 (1989-12-01), Gelfand et al.
patent: 5268289 (1993-12-01), Dahl et al.
Itaya, et al., "Molecular cloning of a ribonuclease H (RNase HI) gene from an extreme thermophile Thermus thermophilus HB8: a thermostable RNase H can functionally replace the Escherichia coli enzyme in vivo", Nucl. Acids Res. 19(16):4443-4449, 1991.
Kanaya, et al., "Expression, Purification, and Characterization of a Recombinant Ribonuclease H from Thermus thermophilus HB8*", J. Biol. Chem. (14):10184-10192, 1992.
Crouch, et al., "Ribonucleases H" in Nuclease (1982) (Levin, S. M. and Roberts, R. J. eds) pp. 211-241, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Kanaya et al., "DNA Sequence of the Gene Coding for Escherichia coli Ribonuclease H," J. Biol. Chem. 258[2]:1276-1281 (1983).
Berkower, I., et al., "Isolation and Characterization of an Endonuclease from Escherichia coli Specific for Ribonucleic Acid in Ribonucleic Acid Deoxyribonucleic Acid Hybrid Structures," J. Biol. Chem. 248[17]:5914-5921 (1973).
Gubler, U., et al., "A simple and very efficient method for generating cDNA libraries," Gene 25:263-269 (1983).
Guatelli, J. C., et al., "Isothermal, in vitro amplication of nucleic acids by a multienzyme reaction modeled after retroviral replication," Proc. Natl. Acad. Sci. USA 87:1874-1878 (1990).
Kuze, et al., "A new vector and RNase H method for the subtractive hybridization," Nucleic Acids Res. 17[2]:807 (1989).
Minshull, J., et al., "The use of single-stranded DNA and RNase H to promote quantitative `hybrid arrest of translation` of mRNA/DNA hybrids in reticulocyte lysate cell-free translations," Nucleic Acids Res. 14[16]:6433-6451 (1986).
Krug, M. S., et al., "Determination of the Molar Concentration of Messenger RNA" Methods in Enzymology 152:262-317 (1987).
Duck, P., et al., "Probe Amplifier System Based on Chimeric Cycling Oligonucleotides," Biotechniques 9[2]:142 (1990).
Crouch, R., in Gene Amplification and Analysis (1981) (Cherikjian, J. G. and Popas, T. S., eds.) 2:217-228, Elsewier, North Holland, New York.
Okayama, H., et al., "High-Efficiency Cloning of Full-Length cDNA," Mol. Cell Biol. 2[2]:161-170 (1982).
Jendrisak, J., "The use of Polyethyleneimine in Protein Purification," Protein Purification: Micro to Macro pp. 75-97 (1987).
Lawyer, F. C., et al., "Isolation, Characterization, and Expression in Escherichia coli of the DNA Polymerase Gene from Thermus aquaticus," J. Biol. Chem. 264[11]:6427-6437 (1989).

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